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See through Walls ! |
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| Maybe not brick walls; but certainly test-tube walls, 96-well plate walls, gel plate walls..... all sorts of plastic and glass walls. There are 2 reasons for the superiority of the Dark Reader compared to UV: |
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| 1. No Blocked Light | ||||||
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| Dark Reader blue light easily passes through transparent materials whereas UV is often blocked or severely attenuated. Consequently, the Dark Reader will be much more effective at exciting fluorophors. | ||||||
| An example of how to block UV: the gel apparatus consists of several layers - the protective outer cover, the buffer reservoir and the electrophoresis plates. Using UV light, absolutely no sample fluorescence is visible. With a Dark Reader Hand Lamp, the EGFP can be clearly seen migrating down the gel. | ||||||
| 2. No Background | ||||||
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| Many plastics fluoresce when exposed to UV light. The result is a high background fluorescence that masks any fluorescence from the actual sample. | ||||||
| An example of background: Fluorescein in a 1.5 mL plastic tube was viewed using either a Dark Reader Hand Lamp or a UV device. In the case of UV irradiation, the fluorescence from the sample is difficult to separate from the fluorescence from the tube itself. Using a Dark Reader, there is essentially no fluorescence from the tube and the fluorescein sample is clearly visible. | ||||||
| The Typical Research Lab | ||||||
| The typical research lab is crammed with apparatus, containers and other devices made from glass or plastics that may severely degrade the usefulness of UV light to detect fluorophors. The data presented in the graph below show that using a Dark Reader instead, results in an 8- to 16-fold improvement in the sensitivity of detection. | ||||||
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| The effectiveness of the Dark Reader for viewing fluorescein. A 2-fold dilution series of fluorescein was variously aliquoted onto assorted laboratory-ware. The dilution series was then viewed by eye on either a Dark Reader transilluminator or a 312 nm UV device and the lowest detectable amount of fluorescein recorded. The actual fluorescein volumes used were: nitrocellulose membrane - 0.5 uL; horizontal gel bed - 2 uL; vertical gel glass plate - 2 uL; 96-well plate - 100 uL; centrifuge tube - 400 uL. |
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